Microscope

Abstract

A microscope includes a movable stage supporting wells arranged in an array, a first imaging unit having a low-magnification objective lens, a second imaging unit having a high-magnification objective lens, a computer determining a representative position of a spheroid based on imaging data of the spheroid acquired by the first imaging unit, and a controller causing respective imaging units to sequentially acquire imaging data of the spheroid in each of the wells. The controller causes the first imaging unit to acquire imaging data for the spheroid in one of the wells, and then causes the stage to adjust the representative position to the optical axis of the high-magnification objective lens, and further causes the second imaging unit to acquire imaging data while causing the first imaging unit to acquire imaging data of the spheroid in another of the wells in synchronization with acquisition by the second imaging unit.

The invention claimed is: 
     
       1. A microscope comprising:
 a movable stage that supports a plurality of sample containers arranged in an array so as to be movable in a horizontal direction; 
 a first objective lens; 
 a second objective lens higher in magnification than the first objective lens, an optical axis of the second objective lens being spaced apart from an optical axis of the first objective lens in an array arrangement direction by a distance substantially equal to an integer multiple of a pitch between two adjacent ones of the sample containers; 
 a first imaging device that acquires, by using the first objective lens, first imaging data of a sample accommodated in each of the sample containers; 
 a second imaging device that acquires, by using the second objective lens, second imaging data of the sample accommodated in each of the sample containers, the second imaging device being provided separately from the first imaging device; 
 a computer configured to determine, for each of the sample containers, a representative position of the sample based on the first imaging data of the sample obtained by the first imaging device; and 
 a controller that controls the first imaging device and the second imaging device to acquire the first and second imaging data of the sample in each of the sample containers by controlling the movable stage to move the sample containers with respect to the first imaging device and the second imaging device, 
 wherein the controller (i) controls the first imaging device to acquire the first imaging data for a first sample in one of the sample containers, (ii) then controls the movable stage to move so as to align the representative position of the first sample determined by the computer based on the first imaging data acquired by the first imaging device with the optical axis of the second objective lens, and (iii) then controls the second imaging device to acquire the second imaging data for the first sample in the one of the sample containers while controlling the first imaging device to acquire the first imaging data of the sample in another of the sample containers in synchronization with the acquisition of the second imaging data for the first sample in the one of the sample containers by the second imaging device. 
 
     
     
       2. The microscope according to  claim 1 , wherein the sample containers are supported by the movable stage in a two-dimensional array of m×n (m>n), and
 the optical axis of the first objective lens and the optical axis of the second objective lens are spaced apart from each other in a direction of m of the sample containers being arrayed. 
 
     
     
       3. The microscope according to  claim 1 , further comprising an irradiator that causes illumination light to penetrate the sample toward the first camera along the optical axis of the first objective lens by irradiating the sample with the illumination light. 
     
     
       4. The microscope according to  claim 1 , further comprising an irradiator that causes illumination light to reflect on the sample toward the first camera along the optical axis of the first objective lens by irradiating the sample with the illumination light. 
     
     
       5. The microscope according to  claim 3 , wherein the sample is irradiated with the illumination light at an angle with respect to the optical axis of the first objective lens, thereby illuminating the sample with focal illumination. 
     
     
       6. The microscope according to  claim 1 , wherein the second objective lens is arranged with the optical axis thereof directed vertically upward. 
     
     
       7. The microscope according to  claim 6 , wherein the first objective lens is arranged with an optical axis thereof directed vertically downward. 
     
     
       8. The microscope according to  claim 1 , wherein the optical axis of the first objective lens and the optical axis of the second objective lens are arranged apart from each other by 4.5 mm, 9 mm, or 18 mm. 
     
     
       9. The microscope according to  claim 1 , further comprising a scanner that scans illumination light on the sample,
 wherein the scanner changes an image magnification by changing a scanning area of the illumination light. 
 
     
     
       10. The microscope according to  claim 1 , wherein:
 the computer is configured to acquire size information of the sample in a plane extending in the horizontal direction based on the first imaging data acquired by the first imaging device, and 
 the controller adjusts an image magnification based on the size information acquired by the computer. 
 
     
     
       11. The microscope according to  claim 1 , further comprising a photodetector that changes a number of imaging pixels. 
     
     
       12. The microscope according to  claim 1 , wherein:
 the computer is configured to (i) acquire size information of the sample in a plane extending in the horizontal direction based on the first imaging data acquired by the first imaging device, and (ii) determine a spherical aberration correction value based on the acquired size information, and 
 the microscope further comprises a correction ring that corrects spherical aberration of the second objective lens according to the spherical aberration correction value determined by the computer. 
 
     
     
       13. The microscope according to  claim 12 , wherein the computer is configured to (i) generate three-dimensional imaging data of the sample based on the second imaging data, and (ii) correct the spherical aberration correction value according to a position of the sample in a vertical direction. 
     
     
       14. The microscope according to  claim 1 , wherein the second objective lens comprises an immersion objective lens. 
     
     
       15. The microscope according to  claim 1 , wherein the first objective lens and the second objective lens are disposed on respective opposite sides of the sample containers arranged in the array. 
     
     
       16. The microscope according to  claim 1 , wherein the another of the sample containers is a sample container from among the sample containers which is adjacent to the one of the sample containers. 
     
     
       17. The microscope according to  claim 1 , wherein the sample is one of a spheroid, a cell, a cellular tissue, and an organoid. 
     
     
       18. The microscope according to  claim 17 , wherein the representative position is a central position of the sample. 
     
     
       19. A non-transitory computer-readable storage medium having a program for controlling a microscope stored thereon, the microscope comprising a movable stage that supports a plurality of sample containers arranged in an array so as to be movable in a horizontal direction, a first objective lens, a second objective lens, a first imaging device, and a second imaging device provided separately from the first imaging device, and the program being executable by a processor of the microscope to control the microscope to perform processes comprising:
 acquiring, by the first imaging device using the first objective lens, first imaging data of a sample accommodated in each of the sample containers; 
 acquiring, by the second imaging device using the second objective lens, second imaging data of the sample accommodated in each of the sample containers, the second objective lens being higher in magnification than the first objective lens, and an optical axis of the second objective lens being spaced apart from an optical axis of the first objective lens in an array arrangement direction by a distance substantially equal to an integer multiple of a pitch between two adjacent ones of the sample containers; 
 determining, for each of the sample containers, a representative position of the sample based on the first imaging data of the sample acquired by the first imaging device; and 
 controlling the first imaging device and the second imaging device to acquire the first and second imaging data of the sample in each of the sample containers by controlling the movable stage to move the sample containers with respect to the first imaging device and the second imaging device, 
 wherein the controlling comprises (i) controlling the first imaging device to acquire the first imaging data for a first sample in one of the sample containers, (ii) then controlling the movable stage to move so as to align the representative position of the first sample determined in the determining based on the first imaging data acquired by the first imaging device with the optical axis of the second objective lens, and (iii) then controlling the second imaging device to acquire the second imaging data for the first sample in the one of the sample containers while controlling the first imaging device to acquire the first imaging data of the sample in another of the sample containers in synchronization with the acquisition of the second imaging data for the first sample in the one of the sample containers by the second imaging device.